Targeting Glioblastoma with a Nuclear-penetrating Anti-DNA Autoantibody

使用核穿透性抗 DNA 自身抗体靶向胶质母细胞瘤

• 批准号: 9912318
• 负责人: James E. Hansen
• 金额: $ 36.64万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9912318
• 关键词: Adenosine; Agonist; Antinuclear Antibodies; Area; Autoantibodies; Binding; Biometry; Biophysics; Blood - brain barrier anatomy; Brain; Cardiotoxicity; Cell Nucleus; Cells; Clinical; Clinical Trials; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; Development; Dose; Double Strand Break Repair; Drug Delivery Systems; Drug Kinetics; Engineering; Environment; Evaluation; Foundations; Genetic; Genetic Heterogeneity; Glioblastoma; Growth; Human; Immunocompetent; Immunodeficient Mouse; Intercellular Junctions; Ligands; Lupus; Malignant Neoplasms; Methods; Modeling; Mus; Mutation; Normal Cell; Nuclear; Nucleoside Transporter; Nucleosides; Operative Surgical Procedures; PTEN gene; Pathway interactions; Patients; Penetration; Pharmaceutical Preparations; Pharmacology; Pilot Projects; Purinergic P1 Receptors; Radiation; Radiation therapy; Radiation-Sensitizing Agents; Relapse; Research Personnel; Risk; Role; Site; Surface; Systemic Lupus Erythematosus; Technology; Testing; Toxic effect; Translating; Work; Xenograft Model; Xenograft procedure; aggressive therapy; blood-brain barrier penetration; blood-brain barrier permeabilization; brain endothelial cell; cancer cell; chemotherapy; drug development; experience; experimental study; extracellular; improved; in vivo; innovation; mortality; mouse model; nanocarrier; neoplastic cell; neuro-oncology; novel; novel strategies; novel therapeutics; restoration; risk minimization; standard of care; stem-like cell; systemic autoimmune disease; tumor; tumor growth; uptake

Glioblastoma multiforme (GBM), the most common primary malignancy of the brain, has high rates of relapse and mortality despite aggressive treatment with surgery, radiation, and chemotherapy. Improved methods to target and treat GBM are needed, but the blood-brain barrier (BBB) and genetic heterogeneity associated with GBM are obstacles to drug development. The autoimmune disease systemic lupus erythematosus offers an unexpected new approach. We discovered that the lupus anti-DNA autoantibody 3E10 penetrates live cell nuclei and inhibits DNA repair in a manner that does not kill normal cells but is toxic to cancer cells with genetic defects in repair of DNA double- strand breaks (DSBs), including PTEN-deficient cancer cells and tumors. 3E10 penetrates cells via a mechanism that requires both extracellular DNA and the ENT2 nucleoside transporter to be present, which facilitates preferential penetration of 3E10 into tumors due to their expression of ENT2 and increased DNA in their environment. ~40% of primary GBM is PTEN-deficient, and we hypothesize that 3E10 will target GBM, have single agent effects against PTEN-deficient GBM, and can serve as a sensitizing agent for radiation therapy and as a drug delivery ligand for GBM regardless of PTEN status. We have re-engineered a 3E10 fragment, hereafter referred to as Deoxymab-1 (DX1), to maximize effect on cancer cells and minimize risks. In preliminary studies DX1 crosses the BBB to localize into and suppress growth of PTEN-deficient GBM in an orthotopic patient-derived xenograft (PDX) mouse model, and delivers conjugated nanocarriers to orthotopic GBM tumors. We now propose studies to help translate DX1 into a novel therapy for GBM. In Aim 1 we pursue studies to elucidate and enhance the mechanism by which DX1 crosses the BBB in GBM. In Aim 2 the effects of DX1, alone or in combination with radiation therapy, on viability and DNA damage accumulation in GBM and normal cells will be determined. DX1 +/- radiation therapy will then be tested in PDX and syngeneic mouse models of GBM to evaluate efficacy and toxicity. In Aim 3 methods to deliver drug-loaded nanocarriers by surface conjugation with DX1 are developed and tested in orthotopic GBM models. We believe use of a modified nuclear-penetrating lupus anti-DNA autoantibody against GBM is an innovative and compelling new strategy that has potential for significant clinical impact, and the proposed studies will establish a foundation for advancing the DX1 technology to clinical trials.

多形性胶质母细胞瘤 (GBM) 是最常见的原发性脑恶性肿瘤，复发率很高 尽管进行了手术、放疗和化疗等积极治疗，但死亡率仍然很高。改进方法 需要针对 GBM 进行靶向治疗，但血脑屏障 (BBB) 和与 GBM 相关的遗传异质性 GBM 是药物开发的障碍。自身免疫性疾病系统性红斑狼疮提供了一种 意想不到的新方法。 我们发现狼疮抗 DNA 自身抗体 3E10 可以穿透活细胞核并抑制 DNA 修复 这种方式不会杀死正常细胞，但对修复 DNA 双链有遗传缺陷的癌细胞具有毒性。 链断裂 (DSB)，包括 PTEN 缺陷的癌细胞和肿瘤。 3E10通过机制穿透细胞 需要细胞外 DNA 和 ENT2 核苷转运蛋白同时存在，这有利于 由于 ENT2 的表达和肿瘤中 DNA 的增加，3E10 优先渗透到肿瘤中 环境。约 40% 的原发性 GBM 缺乏 PTEN，我们假设 3E10 将靶向 GBM，具有 单药对 PTEN 缺陷 GBM 有效，可作为放射治疗和治疗的增敏剂 作为 GBM 的药物递送配体，无论 PTEN 状态如何。 我们重新设计了 3E10 片段，以下称为 Deoxymab-1 (DX1)，以最大限度地提高 癌细胞并最大限度地降低风险。在初步研究中，DX1 穿过 BBB 定位并抑制生长 在原位患者来源的异种移植 (PDX) 小鼠模型中研究 PTEN 缺陷的 GBM，并提供缀合 原位 GBM 肿瘤的纳米载体。我们现在提出研究，帮助将 DX1 转化为一种新疗法 GBM。在目标 1 中，我们进行研究以阐明和增强 DX1 穿过 BBB 的机制 GBM。在目标 2 中，DX1 单独使用或与放射治疗联合使用对活力和 DNA 损伤的影响 将确定 GBM 和正常细胞中的积累。然后将在 PDX 中测试 DX1 +/- 放射治疗 和 GBM 同基因小鼠模型，以评估疗效和毒性。 In Aim 3种载药方法 通过与 DX1 表面缀合的纳米载体在原位 GBM 模型中开发和测试。 我们相信使用改良的核穿透性狼疮抗 DNA 自身抗体来对抗 GBM 是一种创新 以及具有重大临床影响潜力的引人注目的新策略，并且拟议的研究将 为将DX1技术推进临床试验奠定基础。